[Zurück]

M. Lopez-Sanchez, M.J. Ayora-Cañada, A. Molina Díaz, M. Siam, W. Huber, G. Quintas, S. Armenta Estrela, B. Lendl:
"Determination of enzyme activity inhibition by FTIR spectroscopy on the example of fructose bisphosphatase";
Analytical and Bioanalytical Chemistry, 394 (2009), 8; S. 2137 - 2144.

A mid-IR enzymic assay for label-free monitoring of the enzymic reaction of fructose-1,6-bisphosphatase with fructose 1,6-bisphosphate has been proposed. The whole procedure was done in an automated way operating in the stopped flow mode by incorporating a temp.-controlled flow cell in a sequential injection manifold. Fourier transform IR difference spectra were evaluated for kinetic parameters, like the Michaelis-Menten const. (KM) of the enzyme and Vmax of the reaction. The obtained KM of the reaction was 14 3 g L-1 (41 .mu.M). Furthermore, inhibition by AMP (AMP) was evaluated, and the KMApp value was detd. to be 12 2 g L-1 (35 .mu.M) for 7.5 and 15 .mu.M AMP, resp., with Vmax decreasing from 0.1 0.03 to 0.05 0.01 g L-1 min-1. Therefore, AMP exerted a non-competitive inhibition.